PL432230A1 - Method of amplifying complementary DNA in real-time polymerase chain reaction with reverse transcription using gene and region-specific primers for the miRNA-944 precursor - Google Patents

Method of amplifying complementary DNA in real-time polymerase chain reaction with reverse transcription using gene and region-specific primers for the miRNA-944 precursor

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Publication number
PL432230A1
PL432230A1 PL432230A PL43223019A PL432230A1 PL 432230 A1 PL432230 A1 PL 432230A1 PL 432230 A PL432230 A PL 432230A PL 43223019 A PL43223019 A PL 43223019A PL 432230 A1 PL432230 A1 PL 432230A1
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PL
Poland
Prior art keywords
mirna
gene
region
specific primers
cdna
Prior art date
Application number
PL432230A
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Polish (pl)
Other versions
PL237994B1 (en
Inventor
Tomasz Powrózek
Teresa Małecka-Massalska
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Uniwersytet Medyczny W Lublinie
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Publication date
Application filed by Uniwersytet Medyczny W Lublinie filed Critical Uniwersytet Medyczny W Lublinie
Priority to PL432230A priority Critical patent/PL237994B1/en
Publication of PL432230A1 publication Critical patent/PL432230A1/en
Publication of PL237994B1 publication Critical patent/PL237994B1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pathology (AREA)
  • Oncology (AREA)
  • Hospice & Palliative Care (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Przedmiotem wynalazku jest sposób amplifikacji komplementarnego DNA w reakcji łańcuchowej polimerazy w czasie rzeczywistym z odwrotną transkrypcją (RT-qPCR) za pomocą starterów genowo i regiono-specyficznych dla prekursora miRNA-944, który charakteryzuje się tym, że stosuje się startery posiadające następujące sekwencje nukleotydowe: a) startery genowo-specyficzne (służące przepisaniu sekwencji RNA pri-miRNA-944 oraz GAPDH na komplementarne dla nich DNA - cDNA) wykorzystywane w pierwszym etapie RT-qPCR: 1. starter genowo-specyficzny (G1) dla RNA pri-miRNA-944: 5'- ATGGGAGACACAGC - 3', 2. starter genowo-specyficzny (G2) dla RNA GAPDH: 5' - GGACTGAGATTGGC - 3' b) startery regiono-specyficzne (służące amplifikacji cDNA pri-miRNA-944 oraz GAPDH) wykorzystywane w drugim etapie RT-qPCR: 1. starter sensowny (F1) dla cDNA pri-miRNA-944: 5'- GTTCCAGACACATCTCATCTGATA - 3', 2. starter antysensowny (R1) dla cDNA pri-miRNA-944: 5'- TCCCAGACACAGCTCATCCG - 3', 3. starter sensowny (F2) dla cDNA GAPDH: 5,- CCATCTCAGTCGTTCCCAAAGT - 3', 4. starter antysensowny (R2) dla cDNA GAPDH: 5'- AGGTGATCGGTGCTGGTTCC - 3'.The present invention relates to a method of amplifying complementary DNA by real-time reverse transcription polymerase chain reaction (RT-qPCR) with gene and region-specific primers for the miRNA-944 precursor, characterized in that primers having the following nucleotide sequences are used: a) gene-specific primers (for transcribing the RNA sequences of pri-miRNA-944 and GAPDH into complementary DNA - cDNA) used in the first stage of RT-qPCR: 1. gene-specific primer (G1) for pri-miRNA-944 RNA : 5'- ATGGGAGACACAGC - 3 ', 2nd gene-specific primer (G2) for GAPDH RNA: 5' - GGACTGAGATTGGC - 3 'b) region-specific primers (for amplification of pri-miRNA-944 and GAPDH cDNA) used in the second RT-qPCR stage: 1. sense primer (F1) for pri-miRNA-944 cDNA: 5'- GTTCCAGACACATCTCATCTGATA - 3 ', 2.antisense primer (R1) for pri-miRNA-944 cDNA: 5'- TCCCAGACACAGCTCATCCG - 3' 3rd sense primer (F2) for GAPDH cDNA: 5, - CCATCTCAGTCGTTCCCAAAGT - 3 ', 4th antisense primer (R2) for GAPDH cDNA: 5'- AGGTGATCGGTGCTGGTTCC - 3'.

PL432230A 2019-12-16 2019-12-16 Method of amplifying complementary DNA in real-time polymerase chain reaction with reverse transcription using gene and region-specific primers for the miRNA-944 precursor PL237994B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PL432230A PL237994B1 (en) 2019-12-16 2019-12-16 Method of amplifying complementary DNA in real-time polymerase chain reaction with reverse transcription using gene and region-specific primers for the miRNA-944 precursor

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PL432230A PL237994B1 (en) 2019-12-16 2019-12-16 Method of amplifying complementary DNA in real-time polymerase chain reaction with reverse transcription using gene and region-specific primers for the miRNA-944 precursor

Publications (2)

Publication Number Publication Date
PL432230A1 true PL432230A1 (en) 2021-02-22
PL237994B1 PL237994B1 (en) 2021-06-28

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Family Applications (1)

Application Number Title Priority Date Filing Date
PL432230A PL237994B1 (en) 2019-12-16 2019-12-16 Method of amplifying complementary DNA in real-time polymerase chain reaction with reverse transcription using gene and region-specific primers for the miRNA-944 precursor

Country Status (1)

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PL (1) PL237994B1 (en)

Also Published As

Publication number Publication date
PL237994B1 (en) 2021-06-28

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